半乳糖凝集素-1抑制过敏性哮喘小鼠Th2型炎症反应的研究

doi:10.12140/j.issn.1000-7423.2020.02.013

摘要/Abstract

摘要：

目的探讨半乳糖凝集素-1（galectin-1）抑制过敏性哮喘小鼠模型Th2型炎症反应的作用机制。方法 54只BALB/c雌性小鼠随机均分为健康对照组、卵清蛋白（OVA）组和治疗组。在第0、3、7天,OVA组和治疗组小鼠分别皮下注射致敏液[含100 μg OVA混合相同体积10% Al(OH)₃佐剂],健康对照组注射等量生理盐水。最后1次注射后7 d,OVA组和治疗组小鼠分别进行滴鼻激发（50 μg OVA）,健康对照组使用50 μl生理盐水,每天激发1次。治疗组在激发2 h后,给予galectin-1（1 μg/ml）进行滴鼻治疗,健康对照组和OVA组使用生理盐水（10 μl）,连续7 d。第22天,检测3组小鼠气道高反应;收集肺泡灌洗液（BALF）,吉氏染色后,对炎症细胞进行分类并计数;肺组织切片用HE染色后,镜下观察肺组织炎症病理变化;取眼球血,ELISA检测血清中过敏原特异性IgE和Th2型炎症细胞因子白细胞介素-4（IL-4）、IL-5、IL-13,以及γ干扰素（IFN-γ）、IL-10、转化生长因子-β（TGF-β）的表达水平。取脾淋巴细胞,流式细胞术检测调节性T细胞（Treg）的比例。结果气道高反应性结果显示,治疗组小鼠的气道高反应性（Penh值为2.08 ± 0.17）较OVA组（Penh值为5.77 ± 0.64）降低（P < 0.05）。治疗组小鼠BALF中的嗜酸粒细胞、巨噬细胞和中性粒细胞数量分别为（1.33 ± 0.52）×10 ⁴/ml、（1.16 ± 0.41）×10 ⁴/ml、（1.33 ± 0.52）×10 ⁴/ml,与OVA组的[（7.00 ± 1.41）×10 ⁴/ml、（5.00 ± 0.63）×10 ⁴/ml、（5.50 ± 0.84）×10 ⁴/ml]比较,差异有统计学意义（P < 0.05）。肺组织HE染色切片镜检显示,OVA组小鼠支气管周围出现炎症细胞浸润和气道壁增厚;治疗组小鼠的肺部炎症较OVA组改善,且与健康对照组相似。血清ELISA检测结果显示,治疗组血清中过敏原特异性IgE为0.24 ± 0.06,IL-4、IL-5、IL-13的表达水平分别为（104.49 ± 20.76）pg/ml、（82.82 ± 7.71）pg/ml和（31.59 ± 9.78）pg/ml,均较OVA组的[0.87 ± 0.10,（442.72 ± 14.97）、（445.18 ± 35.60）和（434.67 ± 9.78）pg/ml]降低（P < 0.05）。治疗组血清中的IFN-γ、IL-10、TGF-β的表达水平分别为（120.80 ± 9.71）、（63.05 ± 6.05）、（67.89 ± 6.64）pg/ml,均较OVA组的[（47.28 ± 5.01）、（23.89 ± 2.98）、（15.49 ± 3.75）pg/ml]提高（P < 0.05）。流式细胞术检测结果显示,治疗组的Treg比例为（9.64 ± 0.41）%,较OVA组的（1.81 ± 0.48）%增加（P < 0.05）。结论 Galectin-1通过促进机体调节性T细胞和调节性细胞因子IFN-γ、IL-10、TGF-β的生成抑制小鼠过敏性哮喘Th2型炎症反应。

关键词: 过敏性哮喘, 半乳糖凝集素-1, Th2型炎症反应, 调节性T细胞

Abstract:

Objective To investigate the mechanisms underlying the inhibitory effects of galectin-1 on Th2 inflammatory responses in a mouse model of allergic asthma.Methods Fifty-four female BALB/c mice were randomized into control group, ovalbumin (OVA) group, and OVA+galectin-1 treatment group. The mice in the OVA and treatment groups received subcutaneous injections of sensitizer solution [containing 100 μg of OVA in 10% Al(OH)₃] on days 0, 3 and 7, while those in the control group were injected with same volumes of saline. Seven days later, the OVA and treatment groups were challenged by 50 μg of intranasal OVA, while the control group was given 50 μl of saline, once daily for 7 consecutive days. Two hours after each challenge, intranasal galectin-1(1 μg/ml) was applied to the treatment group, while the control and OVA groups were given 10 μl of saline. After completion of treatment on day 22, the airway hyperresponsiveness was examined. Bronchoalveolar lavage fluid (BALF) was collected for Giemsa’s staining to classify and count different inflammatory cells. Lung tissue sections underwent hematoxylin-eosin staining to assess inflammatory pathology. ELISA was performed to analyze protein levels of allergen-specific IgE, Th2 inflammatory cytokines (IL-4, IL-5, and IL-13), IFN-γ, IL-10 and TGF-β in orbital blood serum. Splenic lymphocytes were collected to detect the proportion of regulatory T (Treg) cells by flow cytometry.Results The airway hyperresponsiveness of the OVA+galectin-1 treatment group (Penh value, 2.08 ± 0.17) was significantly lower than that of the OVA group (5.77 ± 0.64) (P < 0.05). The numbers of eosinophils, macrophages and neutrophils in the treatment group were (1.00 ± 0.54) × 10 ⁴/ml, (6.00 ± 0.98) × 10 ⁴/ml and (2.00 ± 0.41) × 10 ⁴/ml, respectively, which were significantly different from those in the OVA group [(3.00 ± 0.04) × 10 ⁴/ml, (4.60 ± 0.55) × 10 ⁴/ml and (2.20 ± 0.30) × 10 ⁴/ml] (P < 0.05). HE staining showed inflammatory cell infiltration and airway wall thickening around the bronchus in the OVA group, while the pulmonary inflammation was significantly ameliorated in the galectin-1+OVA treatment group, decreasing to a level comparable to the control group. ELISA results showed that the serum levels of allergen-specific IgE [(0.24 ± 0.06) pg/ml], IL-4 [(104.49 ± 20.76) pg/ml], IL-5 [(82.82 ± 7.71) pg/ml] and IL-13 [(31.59 ± 9.78) pg/ml] were all significantly reduced compared with those in the OVA group [(0.87 ± 0.10) pg/ml, (442.72 ± 14.97) pg/ml, (445.18 ± 35.60) pg/ml, and (434.67 ± 9.78) pg/ml] (P < 0.05). The protein levels of IFN-γ [(120.80 ± 9.71) pg/ml], IL-10 [(63.05 ± 6.05) pg/ml] and TGF-β [(67.89 ± 6.64) pg/ml] in the galectin-1+OVA group were significantly higher than those in the OVA model group [(47.28 ± 5.01) pg/ml, (63.05 ± 6.05) pg/ml and (15.49 ± 3.75) pg/ml] (P < 0.05). The result of flow cytometry examination indicated that the percentage of Treg cells in the galectin-1+OVA treatment group was (9.64 ± 0.41)%, which was significantly higher than (1.81 ± 0.48)% in the OVA model group. Conclusion Galectin-1 inhibits Th2 inflammatory responses in mice with allergic asthma by promoting productions of Treg cells and regulatory cytokines IFN-γ, IL-10 and TGF-β.

Key words: Allergic asthma, Galectin-1, Th2 inflammatory response, Treg

中图分类号:

R392

智敏, 周粉璇, 官鑫, 钟建文, 罗向前. 半乳糖凝集素-1抑制过敏性哮喘小鼠Th2型炎症反应的研究[J]. 中国寄生虫学与寄生虫病杂志, 2020, 38(2): 213-218.

Min ZHI, Fen-xuan ZHOU, Xin GUAN, Jian-wen ZHONG, Xiang-qian LUO. Galectin-1 inhibits Th2 inflammatory responses in mice with allergic asthma[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2020, 38(2): 213-218.

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